Knockdown of a nutrient amino acid transporter gene LdNAT1 reduces free neutral amino acid contents and impairs Leptinotarsa decemlineata pupation

Fu, Kai‐Yun; Guo, Wen‐Chao; Ahmat, Tursun; Li, Guoqing

doi:10.1038/srep18124

Cited by 27 publications

(23 citation statements)

References 56 publications

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“…The same method as previously reported (Fu et al, 2015) was used to introduce dsRNA into larvae. Newly ecdysed penultimate instar and the last instar larvae were allowed to feed foliage immersed with bacterial suspension containing a dsRNA for 2 days (the penultimate instar larvae) or 3 days (the last instar Figure 6.…”

Section: Dietary Introduction Of Dsrnamentioning

confidence: 99%

Isoform specific roles of Broad‐Complex in larval development in Leptinotarsa decemlineata

Meng

Deng

et al. 2019

Insect Molecular Biology

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Broad‐Complex (BrC) is a downstream target of both 20‐hydroxyecdysone and juvenile hormone signalling. BrC regulates morphogenetic changes between nymphal instars in hemimetabolans, whereas it controls pupal commitment, pupal morphogenesis and inhibits adult differentiation in holometabolans. Among five BrC cDNAs (Z1–Z4 and Z6) identified in the Colorado potato beetle, we found in this work that Z1, Z2 and Z6 were mainly expressed at the last (fourth) instar and prepupal stages, whereas the levels of Z3 and Z4 increased during the penultimate (third) instar stage, peaked at the last instar larval phase and gradually decreased at the prepupal and pupal periods. When knocking down all BrC isoforms by RNA interference (RNAi) at the penultimate instar stage, around 20% of the resultant larvae remained as moribund beetles. These moribund BrC RNAi larvae were completely or partially wrapped in old cuticle. Likewise, a portion of larvae treated for a single double‐stranded RNA of Z3, Z4 or Z6 displayed a degree of similar aberrancies, increasing in the order of isoforms Z6 < Z3 < Z4. When silencing all BrC isoforms at the last instar period, most of the RNAi larvae did not normally pupate or emerge as adults. Separately silencing each of the five zinc finger domains revealed that approximately 70% of the Z1 RNAi larvae remained as prepupae, around 60% of the Z6 RNAi specimens formed aberrant prepupae or pupae and about 60% of the Z2 RNAi beetles became deformed pupae. After removal of the old exuviae, these deformed larvae in which either Z1, Z2 or Z6 was depleted possessed adult prothorax and mesothorax, developing antenna, mouthparts and wing discs. Moreover, less than 50% of the resultant pupae finally emerged as adults when either of Z1, Z2 or Z6 was knocked down. Therefore, our findings reveal, for the first time, that the two roles of BrC in insect groups (ie directing morphogenetic changes during juvenile development and regulating larval–pupal–adult metamorphosis) are played by different BrC isoforms in Leptinotarsa decemlineata.

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Section: Dietary Introduction Of Dsrnamentioning

confidence: 99%

Isoform specific roles of Broad‐Complex in larval development in Leptinotarsa decemlineata

Meng

Deng

et al. 2019

Insect Molecular Biology

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“…For example, the D. melanogaster JhI-21 and minidiscs ( mnd ) are JH-inducible [54]. In Leptinotarsa decemlineata , JH and a JH analog can activate LdNAT1 expression [55]. In this study, the expressions of four genes ( BmNAT1 , BmNAT2 , BmGT1-L , and BmINE ) were up-regulated by JHA (Figure 7).…”

Section: Discussionmentioning

confidence: 62%

Genome-Wide Identification, Characterization and Expression Analysis of the Solute Carrier 6 Gene Family in Silkworm (Bombyx mori)

Tang

Liu

Chen

et al. 2016

IJMS

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The solute carrier 6 (SLC6) gene family, initially known as the neurotransmitter transporters, plays vital roles in the regulation of neurotransmitter signaling, nutrient absorption and motor behavior. In this study, a total of 16 candidate genes were identified as SLC6 family gene homologs in the silkworm (Bombyx mori) genome. Spatio-temporal expression patterns of silkworm SLC6 gene transcripts indicated that these genes were highly and specifically expressed in midgut, brain and gonads; moreover, these genes were expressed primarily at the feeding stage or adult stage. Levels of expression for most midgut-specific and midgut-enriched gene transcripts were down-regulated after starvation but up-regulated after re-feeding. In addition, we observed that expression levels of these genes except for BmSLC6-15 and BmGT1 were markedly up-regulated by a juvenile hormone analog. Moreover, brain-enriched genes showed differential expression patterns during wandering and mating processes, suggesting that these genes may be involved in modulating wandering and mating behaviors. Our results improve our understanding of the expression patterns and potential physiological functions of the SLC6 gene family, and provide valuable information for the comprehensive functional analysis of the SLC6 gene family.

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“…Although our previous results revealed that ingestion of dsRNA can silence target genes in various tissues including neurons, PGs and the corpora allata that producing JH [33–41], we have not compared the RNAi efficacy of dsRNA ingestion with that of dsRNA injection. Here we determined time-effect and concentration-effect curves of two genes, a nutrient amino acid transporter gene LdNAT1 that is mainly expressed in gut [42] and a transient receptor potential cation channel gene LdTrp that is highly transcribed in eyes [43]. Our results showed that injected and fed dsRNAs could reduce approximately 90% of LdNAT1 and LdTrp mRNAs 24 and 36 hours after treatment (S2A and S2B Fig).…”

Section: Resultsmentioning

confidence: 99%

“…In the final larval instar of L . decemlineata , a small 20E rise appears at 4 days after ecdysis [42]. Here, we found that dietary supplement with 20E to generate a premature 20E peak at the foraging stage (Fig 2A), or knockdown of an ecdysteroidogenesis gene (Shade, SHD ) [38] to remove this 20E rise (Fig 2B and S3A Fig), did not affect the phototaxis switch, neither did silencing of a 20E receptor gene EcR (Fig 2C and S3B Fig) or a 20E cascade gene HR3 (Fig 2D and S3C Fig) to eliminate 20E signal.…”

Section: Resultsmentioning

confidence: 99%

Hormonal signaling cascades required for phototaxis switch in wandering Leptinotarsa decemlineata larvae

Meng

Zhu

et al. 2019

PLoS Genet

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Many animals exploit several niches sequentially during their life cycles, a fitness referred to as ontogenetic niche shift (ONS). To successfully accomplish ONS, transition between development stages is often coupled with changes in one or more primitive, instinctive behaviors. Yet, the underlining molecular mechanisms remain elusive. We show here that Leptinotarsa decemlineata larvae finish their ONS at the wandering stage by leaving the plant and pupating in soil. At middle wandering phase, larvae also switch their phototactic behavior, from photophilic at foraging period to photophobic. We find that enhancement of juvenile hormone (JH) signal delays the phototactic switch, and vise verse. Moreover, RNA interference (RNAi)-aided knockdown of LdPTTH (prothoracicotropic hormone gene) or LdTorso (PTTH receptor gene) impairs avoidance response to light, a phenotype nonrescuable by 20-hydroxyecdysone. Consequently, the RNAi beetles pupate at the soil surface or in shallow layer of soil, with most of them failing to construct pupation chambers. Furthermore, a combination of depletion of LdPTTH/LdTorso and disturbance of JH signal causes no additive effects on light avoidance response and pupation site selection. Finally, we establish that TrpA1 (transient receptor potential (TRP) cation channel) is necessary for light avoidance behavior, acting downstream of PTTH. We conclude that JH/PTTH cascade concomitantly regulates metamorphosis and the phototaxis switch, to drive ONS of the wandering beetles from plant into soil to start the immobile pupal stage.

show abstract

Knockdown of a nutrient amino acid transporter gene LdNAT1 reduces free neutral amino acid contents and impairs Leptinotarsa decemlineata pupation

Cited by 27 publications

References 56 publications

Isoform specific roles of Broad‐Complex in larval development in Leptinotarsa decemlineata

Isoform specific roles of Broad‐Complex in larval development in Leptinotarsa decemlineata

Genome-Wide Identification, Characterization and Expression Analysis of the Solute Carrier 6 Gene Family in Silkworm (Bombyx mori)

Hormonal signaling cascades required for phototaxis switch in wandering Leptinotarsa decemlineata larvae

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